摘要
利用高通量测序技术,研究连续传代微生物培养过程中土壤可培养解磷菌的群落变化。采用固体和液体无机磷(IPM)和有机磷培养基(OPM)分别对供试水稻土中解磷菌进行传代富集培养,获得第一代(1st-En)、第二代(2nd-En)和第三代(3rd-En)可培养微生物集合,通过高通量测序技术分析富集物和土壤DNA中细菌16S rRNA基因,评估土壤可培养“潜在”解磷菌群落富集规律及其占本底土著细菌群落的比例。结果表明:供试土壤共检测到本底细菌群落58门、160纲、373目、575科和979属,而利用四种培养基连续传代三次共富集20门、35纲、80目、121科和223属“潜在”解磷菌。OPM富集“潜在”解磷菌分类单元数普遍高于IPM。可培养“潜在”解磷菌在土壤本底土著细菌群落中的比例在门水平最高(8.62%~25.9%),而在其他分类水平上仅为3.22%~12.5%。与本研究构建的已知解磷菌数据库进行比较,土壤本底包括已知解磷菌110属,而四种培养基连续传代三次共富集已知解磷菌83属,对土壤已知解磷菌属的培养比例为75.5%,亦表明土壤中至少约24.5%已知解磷菌属未被富集或遗漏。不同培养基和代次富集了优势解磷菌门,主要包括Proteobacteria、Actinobacteriota和Firmicutes,三者之和为97.20%~99.97%;而在属水平下,不同培养基和代次富集了具有明显生理代谢特征差异的解磷菌属。此外,培养基还富集到大量尚未被证实具有解磷功能或特征基因的细菌属140个,占可培养“潜在”解磷菌群落62.8%,其中绝大部分为数量弱势的稀有属,少数具生长富集优势,如Chelatococcus等。通过将传统微生物培养技术与高通量测序技术相结合,本研究揭示了培养基成分、培养基形态和传代次数均能强烈影响土壤可培养解磷菌多样性评估,为环境解磷菌资源的定向发掘和研究提供了参考。
【Objective】High-throughput sequencing technology was used to study the changes in the diversity of culturable phosphate-solubilizing bacteria(PSB)during serial enrichment,to evaluate the effects of different media and generations on diversity assessment of culturable PSB in paddy soil【.Methods】Through serial passages with inorganic phosphorus medium(IPM)and organic phosphorus medium(OPM)in both solid and liquid forms,culturable“potential”PSB enrichments were obtained from the first(1st-En),second(2nd-En)and third(3rd-En)generation of the media and then for DNA extraction together with the tested soil.Bacterial 16S rRNA genes were analyzed by high-throughput sequencing technology to explore the enrichment laws of soil culturable“potential”PSB and their proportions to soil background indigenous bacterial community.【Results】The results showed that a total of 58 phyla,160 classes,373 orders,575 families and 979 genera were detected in soil background bacterial communities,and 20 phyla,35 orders,80 orders,121 families and 223 genera of the“potential”PSB were cultured by four media after three generations.The diversity of culturable“potential”PSB was generally higher in OPM than IPM and the proportions of culturable“potential”PSB in soil background indigenous bacterial community were the highest at the phylum level(8.62%-25.9%),while only 3.22%-12.5% at other taxonomic levels.Compared with the known PSB database constructed in this study,110 genera of the known PSB existed in soil background indigenous bacterial community;83 genera of the known PSB were existed in the enrichments of four media after three consecutive generations and accounting for 75.5% of the known PSB in the background soil.Thus,at least about 24.5% of the genera of the known PSB in the soil were not enriched or omitted.During the culture of three generations with different media,the culturable dominant PSB mainly included Proteobacteria,Actinobacteriota and Firmicutes,and their relative abundance reached a total of 97
作者
夏围围
张泽霖
邹萌萌
次仁拉姆
陈晓芬
孙祥鑫
王玉芳
贾仲君
XIA Weiwei;ZHANG Zelin;ZOU Mengmeng;Cirenlamu;CHEN Xiaofen;SUN Xiangxin;WANG Yufang;JIA Zhongjun(Jiangsu Key Laboratory of Agricultural Meteorology,School of Ecology and Applied Meteorology,Nanjing University of Information Science and Technology,Nanjing 210044,China;National Engineering&Technology Research Center for Red Soil Improvement,Soil and Fertilizer&Resources and Environment Institute,Jiangxi Academy of Agricultural Sciences,Nanchang 330200,China;State Key Laboratory of Soil and Sustainable Agriculture,Institute of Soil Science,Chinese Academy of Sciences,Nanjing 211135,China;Northeast Institute of Geography and Agroecology,Chinese Academy of Sciences,Changchun 130102,China)
出处
《土壤学报》
CAS
CSCD
北大核心
2024年第6期1680-1693,共14页
Acta Pedologica Sinica
基金
国家自然科学基金项目(42177280)资助。
关键词
解磷菌
无机磷
有机磷
连续传代培养
高通量测序
Phosphate-solubilizing bacteria
Inorganic phosphorus
Organic phosphorus
Serial enrichment
High-throughput sequencing
